Abstract
In this study, we investigated to what extent the stability and transduction capacity of polyplexed DNA can be improved by optimizing the condensing peptide sequence. We have synthesized a small library of cationic peptides, at which the lysine/arginine ratio and the cation charge were varied. All peptides were able to compact DNA, at which polyplexes of short lysine-rich sequences were considerably larger than those of elongated or arginine-rich peptides (GM102 and GM202). In addition, the arginine-rich peptides GM102 and GM202 rendered the polyplexes resistant to plasma incubation or DNase I-mediated digestion. While all peptides were found to improve the transfection efficiency in HepG2 cells, only the GM102- and GM202-derived polyplexes could be specifically targeted to HepG2 cells by incorporation of a ligand-derivatized YKAK8WK peptide. We propose that GM102 and GM202 combine the advantage of small condensing peptides to give small-sized polyplexes with the superior stability of condensing polymers, which makes GM102 and GM202 excellent candidates for future in vivo gene therapy studies.
Similar content being viewed by others
References
Li B et al. Lyophilization of cationic lipid–protamine–DNA (LPD) complexes. J Pharm Sci 2000; 89: 355–364.
Mahato RI . Non-viral peptide-based approaches to gene delivery. J Drug Target 1999; 7: 249–268.
Young JL, Dean DA . Nonviral gene transfer strategies for the vasculature. Microcirculation 2002; 9: 35–49.
Wolschek MF et al. Specific systemic nonviral gene delivery to human hepatocellular carcinoma xenografts in SCID mice. Hepatology 2002; 36: 1106–1114.
Armeanu S et al. Optimization of nonviral gene transfer of vascular smooth muscle cells in vitro and in vivo. Mol Ther 2000; 1: 366–375.
Domashenko A, Gupta S, Cotsarelis G . Efficient delivery of transgenes to human hair follicle progenitor cells using topical lipoplex. Nat Biotechnol 2000; 18: 420–423.
Felgner PL et al. Lipofection: a highly efficient, lipid-mediated DNA-transfection procedure. Proc Natl Acad Sci USA 1987; 84: 7413–7417.
Mahato RI, Kawabata K, Takakura Y, Hashida M . In vivo disposition characteristics of plasmid DNA complexed with cationic liposomes. J Drug Target 1995; 3: 149–157.
Wu CH, Wilson JM, Wu GY . Targeting genes: delivery and persistent expression of a foreign gene driven by mammalian regulatory elements in vivo. J Biol Chem 1989; 264: 16985–16987.
Wilson JM et al. Retrovirus-mediated transduction of adult hepatocytes. Proc Natl Acad Sci USA 1988; 85: 3014–3018.
Wagner E et al. Influenza virus hemagglutinin HA-2 N-terminal fusogenic peptides augment gene transfer by transferrin–polylysine–DNA complexes: toward a synthetic virus-like gene-transfer vehicle. Proc Natl Acad Sci USA 1992; 89: 7934–7938.
Gottschalk S et al. A novel DNA–peptide complex for efficient gene transfer and expression in mammalian cells. Gene Therapy 1996; 3: 48–57.
Van Rossenberg SM et al. Targeted lysosome disruptive elements for improvement of parenchymal liver cell specific gene delivery. J Biol Chem 2002; 277: 45803–45810.
Navarro-Quiroga I et al. Improved neurotensin-vector-mediated gene transfer by the coupling of hemagglutinin HA2 fusogenic peptide and Vp1 SV40 nuclear localization signal. Brain Res Mol Brain Res 2002; 105: 86–97.
Merwin JR et al. Targeted delivery of DNA using YEE(GalNAcAH)3, a synthetic glycopeptide ligand for the asialoglycoprotein receptor. Bioconjug Chem 1994; 5: 612–620.
Schaffer DV, Lauffenburger DA . Targeted synthetic gene delivery vectors. Curr Opin Mol Ther 2000; 2: 155–161.
Kircheis R et al. Coupling of cell-binding ligands to polyethylenimine for targeted gene delivery. Gene Therapy 1997; 4: 409–418.
Boussif O et al. A versatile vector for gene and oligonucleotide transfer into cells in culture and in vivo: polyethylenimine. Proc Natl Acad Sci USA 1995; 92: 7297–7301.
Guillem VM et al. Targeted oligonucleotide delivery in human lymphoma cell lines using a polyethyleneimine based immunopolyplex. J Control Release 2002; 83: 133–146.
Haensler J, Szoka Jr FC . Polyamidoamine cascade polymers mediate efficient transfection of cells in culture. Bioconjug Chem 1993; 4: 372–379.
Plank C, Mechtler K, Szoka Jr FC, Wagner E . Activation of the complement system by synthetic DNA complexes: a potential barrier for intravenous gene delivery. Hum Gene Ther 1996; 7: 1437–1446.
Sparrow JT et al. Synthetic peptide-based DNA complexes for nonviral gene delivery. Adv Drug Deliv Rev 1998; 30: 115–131.
McKenzie DL, Collard WT, Rice KG . Comparative gene transfer efficiency of low molecular weight polylysine DNA-condensing peptides. J Pept Res 1999; 54: 311–318.
Wadhwa MS et al. Peptide-mediated gene delivery: influence of peptide structure on gene expression. Bioconjug Chem 1997; 8: 81–88.
Niidome T et al. Gene transfer into hepatoma cells mediated by galactose-modified alpha-helical peptides. Biomaterials 2000; 21: 1811–1819.
Wadhwa MS, Knoell DL, Young AP, Rice KG . Targeted gene delivery with a low molecular weight glycopeptide carrier. Bioconjug Chem 1995; 6: 283–291.
Zou Y, Zong G, Ling YH, Perez-Soler R . Development of cationic liposome formulations for intratracheal gene therapy of early lung cancer. Cancer Gene Ther 2000; 7: 683–696.
Wolfert MA, Seymour LW . Atomic force microscopic analysis of the influence of the molecular weight of poly(L)lysine on the size of polyelectrolyte complexes formed with DNA. Gene Therapy 1996; 3: 269–273.
Lacey Jr JC, Pruitt KM . Drug–biomolecule interactions: interactions of mononucleotides and polybasic amino acids. J Pharm Sci 1975; 64: 473–477.
Plank C, Tang MX, Wolfe AR, Szoka Jr FC . Branched cationic peptides for gene delivery: role of type and number of cationic residues in formation and in vitro activity of DNA polyplexes. Hum Gene Ther 1999; 10: 319–332.
Dash PR et al. Factors affecting blood clearance and in vivo distribution of polyelectrolyte complexes for gene delivery. Gene Therapy 1999; 6: 643–650.
Kalderon D, Roberts BL, Richardson WD, Smith AE . A short amino acid sequence able to specify nuclear location. Cell 1984; 39: 499–509.
Lanford RE, Kanda P, Kennedy RC . Induction of nuclear transport with a synthetic peptide homologous to the SV40 T antigen transport signal. Cell 1986; 46: 575–582.
Perales JC et al. Biochemical and functional characterization of DNA complexes capable of targeting genes to hepatocytes via the asialoglycoprotein receptor. J Biol Chem 1997; 272: 7398–7407.
Chiou HC et al. Enhanced resistance to nuclease degradation of nucleic acids complexed to asialoglycoprotein–polylysine carriers. Nucleic Acids Res 1994; 22: 5439–5446.
Cristiano RJ, Smith LC, Woo SL . Hepatic gene therapy: adenovirus enhancement of receptor-mediated gene delivery and expression in primary hepatocytes. Proc Natl Acad Sci USA 1993; 90: 2122–2126.
Oh YK et al. Polyethylenimine-mediated cellular uptake, nucleus trafficking and expression of cytokine plasmid DNA. Gene Therapy 2002; 9: 1627–1632.
Gausepohl H, Boulin C, Kraft M, Frank RW . Automated multiple peptide synthesis. Pept Res 1992; 5: 315–320.
Sliedregt LA et al. Design and synthesis of a multivalent homing device for targeting to murine CD22. Bioorg Med Chem 2001; 9: 85–97.
Commerford SL . In vitro iodination of nucleic acids. Methods Enzymol 1980; 70: 247–252.
Cotten M, Wagner E, Birnstiel ML . Receptor-mediated transport of DNA into eukaryotic cells. Methods Enzymol 1993; 217: 217618–217644.
Acknowledgements
This study was supported by the Netherlands Foundation for Scientific Research (NWO, project 901-01-096), the Netherlands Heart Foundation (project M93 001) and the Chemical Sciences/Foundation of Technical Sciences (CW/STW, project 349-4779).
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
van Rossenberg, S., van Keulen, A., Drijfhout, JW. et al. Stable polyplexes based on arginine-containing oligopeptides for in vivo gene delivery. Gene Ther 11, 457–464 (2004). https://doi.org/10.1038/sj.gt.3302183
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1038/sj.gt.3302183
- Springer Nature Limited